Cervical cancer, the second most prevalent cancer of women worldwide, and the principal cancer of women in most developing countries, has been clearly linked to infection by human papillomavirus (HPV). In virtually all cervical cancer, HPV DNA is found integrated into the host genome, which results in the increased production of viral transforming proteins. In contrast to the morphologically-based traditional Pap test, molecular diagnostic methods detect HPV DNAs in cervical tissue, identifying whether a cancer-linked viral type is present. However, this diagnostic approach has not reduced morbidity and mortality, nor improved disease staging. Recent detection methods have focused on detection of HPV RNA using reverse transcriptase polymerase chain reaction. Although sensitive, no cytological examination of the cell sample is possible and the method continues to be plagued by procedural problems. In situ hybridization allows direct visualization of viral RNAs in addition to cytological examination; however, current in situ hybridization methods are time consuming, difficult to perform, and consist of many procedural steps. This Small Business Innovation Research project proposes to develop a rapid fluorescence in situ hybridization assay for detecting HPV RNAs. [unreadable] [unreadable]